FIGURE 6.
RPA-sedDNA complexes accumulate when gap-filling DNA polymerases are inhibited with aphidicolin. A, quiescent U2OS cells were left untreated or were pretreated for 30 min with 1 μg/ml aphidicolin (APH) prior to exposure to 10 J/m2 of UV. At the indicated time points, genomic DNA was isolated from the cells and analyzed by immunoslot blotting with an anti-(6-4)PP antibody. B, genomic DNA from UV-irradiated cells was analyzed as in A except that an anti-CPD antibody was used for immunoblotting. C, whole cell lysates were prepared from quiescent U2OS cells with the indicated treatments and then subjected to immunoprecipitation with an anti-RPA34 (RPA) antibody. Protein content was analyzed by immunoblotting, and total sedDNA content was analyzed by radiolabeling, urea-PAGE, and phosphorimager analysis. Results from three independent experiments (average and S.D.) are graphed.